Composition for pulmonary administration comprising a drug and a hydrophobic amino acid

ABSTRACT

According to the subject invention, dispersible dry powder pharmaceutical-based compositions are provided, including methods for their manufacture and dry powder dispersion devices. A dispersible dry powder pharmaceutical-based composition is one having a moisture content of less than about 10% by weight (% w) water, usually below about 5% w and preferably less than about 3% w; a particle size of about 1.0-5.0 μm mass median diameter (MMD), usually 1.0-4.0 μm MMD, and preferably 1.0-3.0 μm MMD; a delivered dose of about &gt;30%, usually &gt;40%, preferably &gt;50%, and most preferred &gt;60%; and an aerosol particle size distribution of about 1.0-5.0 μm mass median aerodynamic diameter (MMAD), usually 1.5-4.5 μm MMAD, and preferably 1.5-4.0 μm MMAD. Such compositions are of pharmaceutical grade purity.

[0001] This application is a Continuation of U.S. patent applicationSer. No. 08/423,515, filed on Apr. 14, 1995 and is aContinuation-in-Part of the following U.S. patent applications Ser. No.07/910,048, filed Jul. 8, 1992, now U.S. Pat. No. 4,458,135; Ser. No.08/417,507, filed Apr. 4, 1995, now abandoned, which is a file wrappercontinuation of Ser. No. 08/044,358, filed Apr. 7, 1993, now abandoned;Ser. No. 08/232,849, filed Apr. 25, 1994, now U.S. Pat. No. 5,607,915;Ser. No. 08/309,691, filed Sep. 21, 1994, now U.S. Pat. No. 5,785,049;Ser. No. 08/246,034, filed May 18, 1994, now abandoned; Ser. No.08/313,707, filed Sep. 27, 1994, now abandoned; and Ser. No. 08/383,475,filed Feb. 1, 1995, the full disclosures of which are incorporatedherein by reference.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates generally to methods andcompositions for the dry powder formulation of pharmaceuticals,including macromolecules, for pulmonary delivery.

[0004] Over the years, certain drugs have been sold in compositionssuitable for forming a drug dispersion for oral inhalation (pulmonarydelivery) to treat various conditions in humans. Such pulmonary drugdelivery compositions are designed to be delivered by inhalation by thepatient of a drug dispersion so that the active drug within thedispersion can reach the lung. It has been found that certain drugsdelivered to the lung are readily absorbed through the alveolar regiondirectly into blood circulation. Pulmonary delivery is particularlypromising for the delivery of macromolecules (proteins, polypeptides andnucleic acids) which are difficult to deliver by other routes ofadministration. Such pulmonary delivery can be effective both forsystemic delivery and for localized delivery to treat diseases of thelungs.

[0005] Pulmonary drug delivery can itself be achieved by differentapproaches, including liquid nebulizers, aerosol-based metered doseinhalers (MDI's), and dry powder dispersion devices. Aerosol-based MDI'sare losing favor because they rely on the use of chlorofluorocarbons(CFC's), which are being banned because of their adverse effect on theozone layer. Dry powder dispersion devices, which do not rely on CFCaerosol technology, are promising for delivering drugs that may bereadily formulated as dry powders. Many otherwise labile macromoleculesmay be stably stored as lyophilized or spray-dried powders by themselvesor in combination with suitable powder carriers. The ability to deliverpharmaceutical compositions as dry powders, however, is problematic incertain respects. The dosage of many pharmaceutical compositions isoften critical so it is necessary that any dry powder delivery system beable to accurately, precisely, and reliably deliver the intended amountof drug. Moreover, many pharmaceutical compositions are quite expensive.Thus, the ability to efficiently deliver the dry powders with a minimalloss of drug is critical. It is also essential that the powder bereadily dispersible prior to inhalation by the patient in order toassure adequate distribution and systemic absorption.

[0006] A particularly promising approach for the pulmonary delivery ofdry powder drugs utilizes a hand-held device with a hand pump forproviding a source of pressurized gas. The pressurized gas is abruptlyreleased through a powder dispersion device, such as a venturi nozzle,and the dispersed powder made available for patient inhalation. Whileadvantageous in many respects, such hand-held devices are problematic ina number of other respects. The particles being delivered are less than10 μm in size, usually in the range from 1 μm to 5 μm, making powderhandling and dispersion more difficult than with larger particles. Theproblems are exacerbated by the relatively small volumes of pressurizedgas, which are available using hand-actuated pumps. In particular,venturi dispersion devices are unsuitable for difficult-to-dispersepowders when only small volumes of pressurized gas are available.Another requirement for hand-held and other powder delivery devices isefficiency. It is important that the concentration of drug in the bolusof gas be relatively high to reduce the number of breaths required toachieve a total dosage. The ability to achieve both adequate dispersionand small dispersed volumes is a significant technical challenge thatrequires in part that each unit dosage of the powdered composition bereadily and reliably dispersible.

[0007] 2. Description of the Relevant Literature

[0008] Dry powder dispersion devices for medicaments are described in anumber of patent documents. U.S. Pat. No. 3,921,637 describes a manualpump with needles for piercing through a single capsule of powderedmedicine. The use of multiple receptacle disks or strips of medicationis described in European Patent Application No. EP 0 467 172 (where areciprocatable punch is used to open a blister pack); InternationalPatent Publication Nos. WO 91/02558; WO 93/09832; U.S. Pat. Nos.4,627,432; 4,811,731; 5,035,237; 5,048,514; 4,446,862; 5,048,514; and4,446,862. Other patents which show puncturing of single medicationcapsules include U.S. Pat. Nos. 4,338,931; 3,991,761; 4,249,526;4,069,819; 4,995,385; 4,889,114; and 4,884,565; and European PatentApplication No. EP 469 814. International Patent Publication No. WO90/07351 describes a hand-held pump device with a loose powderreservoir.

[0009] A dry powder sonic velocity disperser is described in Witham andGates, Dry Dispersion with Sonic Velocity Nozzles, presented at theworkshop on Dissemination Techniques for Smoke and Obscurants, ChemicalSystems Laboratory, Aberdeen Proving Ground, Md., Mar. 14-16, 1983.

[0010] U.S. Pat. Nos. 4,926,852 and 4,790,305, describe a type of“spacer” for use with a metered dose inhaler. The spacer defines a largecylindrical volume which receives an axially directed burst of drug froma propellant-driven drug supply. U.S. Pat. No. 5,027,806 is animprovement over the '852 and '305 patents, having a conical holdingchamber which receives an axial burst of drug. U.S. Pat. No. 4,624,251,describes a nebulizer connected to a mixing chamber to permit acontinuous recycling of gas through the nebulizer. U.S. Pat. No.4,677,975 is described above. European Patent Application No. 0 347 779describes an expandable spacer for a metered dose inhaler having aone-way valve on the mouthpiece. International Patent Publication No. WO90/07351 describes a dry powder oral inhaler having a pressurized gassource (a piston pump) which draws a measured amount of powder into aventuri arrangement.

[0011] The respiratory delivery of aerosolized aqueous insulin solutionsis described in a number of references, beginning with Gansslen (1925)Klin. Wochenschr. 4:71 and including Laube et al. (1993) JAMA269:2106-21-9; Elliott et al. (1987) Aust. Paediatr. J. 23:293-297;Wigley et al. (1971) Diabetes 20:552-556. Corthorpe et al. (1992) PharmRes 9:764-768; Govinda (1959) Indian J. Physiol. Pharmacol. 3:161-167;Hastings et al. (1992) J. Appl. Physiol. 73:1310-1316; Liu et al. (1993)JAMA 269:2106-2109; Nagano et al. (1985) Jikeikai Med. J. 32:503-506;Sakr (1992) Int. J. Phar. 86:1-7; and Yoshida et al. (1987) Clin. Res.35:160-166. Pulmonary delivery of dry powder medicaments, such asinsulin, in a large particle carrier vehicle is described in U.S. Pat.No. 5,254,330. A metered dose inhaler (MDI) for delivering crystallineinsulin suspended in a propellant is described in Lee and Sciara (1976)J. Pharm. Sci. 65:567-572. A MDI for delivering insulin into a spacerfor regulating inhalation flow rate is described in U.S. Pat. No.5,320,094. The intrabronchial administration of recombinant insulin isbriefly described in Schluter et al. (Abstract) (1984) Diabetes 33:75Aand Köhler et al. (1987) Atemw. Lungenkrkh. 13:230-232. Intranasal andrespiratory delivery of a variety of polypeptides, including insulin, inthe presence of an enhancer, are described in U.S. Pat. No. 5,011,678and Nagai et al. (1984) J. Contr. Rel. 1:15-22. Intranasal delivery ofinsulin in the presence of enhancers and/or contained in controlledrelease formulations are described in U.S. Pat. Nos. 5,204,108;4,294,829; and 4,153,689; International Patent Publication Nos. WO93/02712, WO 91/02545, WO 90/09780, and WO 88/04556; Great BritainPatent No. 1,527,605; Ryden and Edman (1992) Int. J. Pharm. 83:1-10; andBjork and Edman (1988) Int. J. Pharm. 47:233-238. The preparation andstability of amorphous insulin were described by Rigsbee and Pikal atthe American Association of Pharmaceutical Sciences (AAPS), Nov. 14-18,1993, Lake Buena Vista, Fla. Methods for spray drying polypeptide,polynucleotide and other labile drugs in a carrier which forms anamorphous structure which stabilizes the drug are described in EuropeanPatent Application No. 520 748. (AAPS), Nov. 14-18, 1993, Lake BuenaVista, Fla.

[0012] Stribling et al. (1992) J. Biopharm. Sci. 3:255-263, describesthe aerosol delivery of plasmids carrying a chloramphenicolacetyltransferase (CAT) reporter gene to mice. The plasmids wereincorporated in DOTMA or cholesterol liposomes, and aqueous suspensionsof the liposomes were nebulized into a small animal aerosol deliverychamber. Mice breathing the aerosol were found to at least transientlyexpress CAT activity in their lung cells. Rosenfeld et al. (1991)Science: 252:431-434, describes the in vivo delivery of an alpha-1antitrypsin gene to rats, with secretion of the gene product beingobservable for at least one week. The gene was diluted in saline andinstilled directly into the rat trachea. Friedman (1989) Science244:1275-1281 is a review article describing human gene therapystrategies.

[0013] U.S. Pat. Nos. 4,833,125 and 4,698,328, describe theadministration of active parathyroid hormone fragments in combinationwith vitamin D or a dietary calcium supplement. Suggested administrationroutes include parenteral by injection, rapid infusion, nasopharyngealabsorption, dermal absorption, or oral. See also, Neer et al. (1987)Osteoporosis 53:829-835. U.S. Pat. No. 5,011,678, describes the use ofamphophilic steroids as a penetration enhancer for nasal orbronchopulmonary delivery of proteins and polypeptides, listingparathyroid hormone as one of a “veritable host” of proteins which couldbe delivered with the enhancer. Parathyroid hormone (full length) issecreted naturally from the parathyroid gland as a series of spikes in apulsatile fashion which is analogous to pituitary hormones (Harms et al.(1987) Int. Symp. on Osteoporosis, Aalborg, Abstract 232). The fulllength hormone is rapidly broken down in the circulation into severalfragments which are the dominant serum forms. It is hypothesized that anintermittent or pulsatile secretion pattern for parathyroid hormone isnecessary to maintain its bone restoring properties (Hesch et al. (1988)Calcif. Tissue Int. 42:341-344 and Habener et al. (1971). Proc. Natl.Acad. Sci USA 68:2986-2991). Patton and Platz (1992) Adv. Drug Deliver.Rev. 8:179-196, describe methods for delivering proteins andpolypeptides by inhalation through the deep lung.

[0014] The aerosolization of protein therapeutic agents, includingalpha-1 antitrypsin, is disclosed in European Patent Application No. EP0 289 336. The use of alpha-1 antityrpsin for treating pulmonaryinflammation is disclosed in U.S. Pat. No. 5,093,316.

[0015] Therapeutic aerosol formulations, including calcitonin, aredisclosed in International Patent Publication No. WO 90/09781.

[0016] Methods and compositions for inhibiting neutrophil elastase andcathespin G employing aerosolized 2-0-desulfated heparin are disclosedin International Patent Publication No. WO 94/02107.

[0017] Interleukin-1 receptor compositions are disclosed in U.S. Pat.Nos. 4,968,607, 5,081,228 and 5,180,812.

[0018] Aerosol formulations of interferons have been produced forpulmonary delivery as described in International Patent Publication No.WO 91/16038. International Patent Publication No. WO 91/16038 teachesadding a surfactant or the like to improve the dispersibility of a humaninterferon from a CFC delivery system. Methods and compositions for thepreparation of solid polypeptide microparticles as a pharmaceuticalaerosol formulation are disclosed in International Patent PublicationNo. WO 91/16038. The purification of proteins of molecular weight inexcess of 12,000, including human IFN is disclosed in U.S. Pat. No.4,503,035. Low pH pharmaceutical compositions of recombinant IFN-betaare disclosed in International Patent Publication No. WO 89/05158.

[0019] 3. Objects of the Invention

[0020] An object of the present invention is to provide a pharmaceuticalcomposition suitable for long-term pulmonary administration to a patientin need thereof.

[0021] Another object of this invention is to provide apharmaceutical-containing dispersible dry powdered composition that isadministered by inhalation in a manner that is free of a liquidpropellant such as a CFC, HFC or carbon dioxide.

[0022] Another object of this invention is to provide apharmaceutical-containing dispersible dry powdered composition that canbe easily manufactured by a method that maintains a high percentage ofpharmaceutical activity.

[0023] Another object of this invention is to provide a manufacturablemethod for the production of a pharmaceutical composition of sufficientpurity.

[0024] Still another object of this invention is to provide apharmaceutical-containing dispersible dry powdered composition thatexhibits a high level of stability.

[0025] Other objects may be apparent to one of ordinary skill uponreviewing the following specification and claims.

SUMMARY OF THE INVENTION

[0026] According to the subject invention, dispersible dry powderpharmaceutical-based compositions are provided, including methods fortheir manufacture and dry powder dispersion devices. A dispersible drypowder pharmaceutical-based composition is one having a moisture contentof less than about 10% by weight (% w) water, usually below about 5% wand preferably less than about 3% w; a particle size of about 1.0-5.0 μmmass median diameter (MMD), usually 1.0-4.0 μm MMD, and preferably1.0-3.0 μm MMD; a delivered dose of about >30%, usually >40%,preferably >50%, and most preferred >60%; and an aerosol particle sizedistribution of about 1.0-5.0 μm mass median aerodynamic diameter(MMAD), usually 1.5-4.5 μm MMAD, and preferably 1.5-4.0 MMAD. Suchcompositions are of pharmaceutical grade purity.

DESCRIPTION OF SPECIFIC EMBODIMENTS

[0027] The present invention is based at least in part on thedispersibility characteristics of the pharmaceutical-based dry powdercompositions produced according to the present invention. Thedispersibility characteristics of the subject pharmaceutical-basedcompositions means that they are more suitable for use in pulmonarydelivery devices than compositions prepared by other methods. Thecompositions of the invention are readily aerosolized and rapidlyabsorbed through the lungs of a host when delivered by a dry powderinhaler.

DEFINITIONS

[0028] In interpreting the claims to the various aspects of thisinvention, there are several important definitions that should beconsidered.

[0029] The term “dispersibility” or “dispersible” means a dry powderhaving a moisture content of less than about 10% by weight (% w) water,usually below about 5% w and preferably less than about 3% w; a particlesize of about 1.0-5.0 μm mass median diameter (MMD), usually 1.0-4.0 μmMMD, and preferably 1.0-3.0 μm MMD; a delivered dose of about >30%,usually >40%, preferably >50%, and most preferred >60%; and an aerosolparticle size distribution of about 1.0-5.0 μm mass median aerodynamicdiameter (MMAD), usually 1.5-4.5 μm MMAD, and preferably 1.5-4.0 μmMMAD. Methods and compositions for improving dispersibility aredisclosed in U.S. application Ser. No. 08/423,568, filed Apr. 14, 1995,the disclosure of which is hereby incorporated by reference.

[0030] The term “powder” means a composition that consists of finelydispersed solid particles that are free flowing and capable of beingreadily dispersed in an inhalation device and subsequently inhaled by asubject so that the particles reach the lungs to permit penetration intothe alveoli. Thus, the powder is said to be “respirable.” Preferably theaverage particle size is less than about 10 microns (μm) in diameterwith a relatively uniform spheroidal shape distribution. More preferablythe diameter is less than about 7.5 μm and most preferably less thanabout 5.0 μm. Usually the particle size distribution is between about0.1 μm and about 5 μm in diameter, particularly about 0.3 μm to about 5μm.

[0031] The term “dry” means that the composition has a moisture contentsuch that the particles are readily dispersible in an inhalation deviceto form an aerosol. This moisture content is generally below about 10%by weight (% w) water, usually below about 5% w and preferably less thanabout 3% w.

[0032] The term “therapeutically effective amount” is the amount presentin the composition that is needed to provide the desired level of drugin the subject to be treated to give the anticipated physiologicalresponse. This amount is determined for each drug on a case-by-casebasis. Guidelines are given hereafter.

[0033] The term “physiologically effective amount” is that amountdelivered to a subject to give the desired palliative or curativeeffect. This amount is specific for each drug and its ultimate approveddosage level. Guidelines are given hereafter.

[0034] The term “pharmaceutically acceptable carrier” means that thecarrier can be taken into the lungs with no significant adversetoxicological effects on the lungs.

COMPOSITIONS OF THE INVENTION

[0035] One aspect of this invention is a dispersiblepharmaceutical-based dry powder composition for pulmonary delivery, thecomposition comprising a therapeutically effective amount of apharmaceutical in combination with a pharmaceutically acceptablecarrier.

[0036] In general, the compositions of this invention are suitable forpulmonary delivery because of their dispersibility characteristics. Suchcompositions were not previously known in the art. In the dry state, thepharmaceutical may be in crystalline or amorphous form. Some examples ofpharmaceutical compositions suitable for formulation into dispersibledry powders are listed in Table 1. These include macromolecule andnon-macromolecule-based pharmaceuticals, usually macromolecules, withinsulin, interleukin-1 receptor, parathyroid hormone (PTH-34), alpha-1antitrypsin, calcitonin, low molecular weight heparin, heparin,interferon, and nucleic acids being preferred.

[0037] A therapeutically effective amount of active pharmaceutical willvary in the composition depending on the biological activity of the drugemployed and the amount needed in a unit dosage form. Because thesubject compounds are dispersible, it is highly preferred that they bemanufactured in a unit dosage form in a manner that allows for readymanipulation by the formulator and by the consumer. This generally meansthat a unit dosage will be between about 0.5 mg and 15 mg of totalmaterial in the dry powder composition, preferably between about 2 mgand 10 mg. Generally, the amount of drug in the composition will varyfrom about 0.05% w to about 99.0% w. Most preferably the compositionwill be about 0.2% to about 97.0% w drug.

[0038] The amount of the pharmaceutically acceptable carrier is thatamount needed to provide the necessary stability, dispersibility,consistency and bulking characteristics to ensure a uniform pulmonarydelivery of the composition to a subject in need thereof. Numericallythe amount may be from about 0.05% w to about 99.95% w, depending on theactivity of the drug being employed. Preferably about 5% w to about 95%w will be used.

[0039] The carrier may be one or a combination of two or morepharmaceutical excipients, but will generally be substantially free ofany “penetration enhancers.” Penetration enhancers are surface activecompounds which promote penetration of a drug through a mucosal membraneor lining and are proposed for use in intranasal, intrarectal, andintravaginal drug formulations. Exemplary penetration enhancers includebile salts, e.g., taurocholate, glycocholate, and deoxycholate;fusidates, e.g., taurodehydrofusidate; and biocompatible detergents,e.g., Tweens, Laureth-9, and the like. The use of penetration enhancersin formulations for the lungs, however, is generally undesirable becausethe epithelial blood barrier in the lung can be adversely affected bysuch surface active compounds. The dry powder compositions of thepresent invention are readily absorbed in the lungs without the need toemploy penetration enhancers.

[0040] The types of pharmaceutical excipients that are useful ascarriers in this invention include stabilizers such as human serumalbumin (HSA), bulking agents such as carbohydrates, amino acids andpolypeptides; pH adjusters or buffers; salts such as sodium chloride;and the like. These carriers may be in a crystalline or amorphous formor may be a mixture of the two.

[0041] It has been found that HSA is particularly valuable as a carrierin that it provides improved dispersibility.

[0042] Bulking agents that are particularly valuable include compatiblecarbohydrates, polypeptides, amino acids or combinations thereof.Suitable carbohydrates include monosaccharides such as galactose,D-mannose, sorbose, and the like; disaccharides, such as lactose,trehalose, and the like; cyclodextrins, such as2-hydroxypropyl-β-cyclodextrin; and polysaccharides, such as raffinose,maltodextrins, dextrans, and the like; alditols, such as mannitol,xylitol, and the like. A preferred group of carbohydrates includeslactose, trehalose, raffinose, maltodextrins, and mannitol. Suitablepolypeptides include aspartame. Amino acids include alanine and glycine,with glycine being preferred.

[0043] Additives, which are minor components of the composition of thisinvention, may be included for conformational stability during spraydrying and for improving dispersibility of the powder. These additivesinclude hydrophobic amino acids such as tryptophan, tyrosine, leucine,phenylalanine, and the like.

[0044] Suitable pH adjusters or buffers include organic salts preparedfrom organic acids and bases, such as sodium citrate, sodium ascorbate,and the like; sodium citrate is preferred.

[0045] The unit dosage form, method of treatment, and process ofpreparation of this invention are described hereafter.

[0046] Unit Dosage Form.

[0047] Another aspect of this invention is a unit dosage form forpulmonary delivery of dispersible dry powder pharmaceutical-basedcompositions, which dosage form comprises a unit dosage receptaclecontaining a pharmaceutical-based dry powder composition, whichcomposition comprises a therapeutically effective amount of apharmaceutical in combination with a pharmaceutically acceptablecarrier.

[0048] In this aspect of the invention, the composition of thisinvention (as discussed hereinbefore) is placed within a suitable dosagereceptacle in an amount sufficient to provide a subject with drug for aunit dosage treatment. The dosage receptacle is one that fits within asuitable inhalation device to allow for the aerosolization of theinterferon-based dry powder composition by dispersion into a gas streamto form an aerosol and then capturing the aerosol so produced in achamber having a mouthpiece attached for subsequent inhalation by asubject in need of treatment. Such a dosage receptacle includes anycontainer enclosing the composition known in the art such as gelatin orplastic capsules with a removable portion that allows a stream of gas(e.g., air) to be directed into the container to disperse the dry powdercomposition. Such containers are exemplified by those shown in U.S. Pat.Nos. 4,227,522 issued Oct. 14, 1980; 4,192,309 issued Mar. 11, 1980; and4,105,027 issued Aug. 8, 1978. Suitable containers also include thoseused in conjunction with Glaxo's Ventolin® Rotohaler brand powderinhaler or Fison's Spinhaler® brand powder inhaler. Another suitableunit-dose container which provides a superior moisture barrier is formedfrom an aluminum foil plastic laminate. The pharmaceutical-based powderis filled by weight or by volume into the depression in the formablefoil and hermetically sealed with a covering foil-plastic laminate. Sucha container for use with a powder inhalation device is described in U.S.Pat. No. 4,778,054 and is used with Glaxo's Diskhaler® (U.S. Pat. Nos.4,627,432; 4,811,731; and 5,035,237). All of these references areincorporated herein by reference.

[0049] Method of Treating a Disease State.

[0050] Another aspect of this invention is a method of treating acondition responsive to treatment by a pharmaceutical of interest, whichmethod comprises pulmonarily administering to a subject in need thereofa physiologically effective amount of a dispersible pharmaceutical-baseddry powder composition that comprises a therapeutically effective amountof drug in combination with a pharmaceutically acceptable carrier.

[0051] Conditions that may be treated by the compositions of this aredescribed in Table 1.

[0052] The physiologically effective amount needed to treat a particularcondition or disease state will depend on the individual, the condition,length of treatment, the regularity of treatment, the type of drug, andother factors, but can be determined by one of ordinary skill in themedicinal arts.

[0053] It is presently believed that the effective absorption by a hostof dry powder composition according to the present invention resultsfrom a rapid dissolution in the ultra-thin (<0.1 μm) fluid layer of thealveolar lining of the lung. The particles of the present invention thushave a mean size which is from 10 to 50 times larger than the lung fluidlayer, making it unexpected that the particles are dissolved and theinterferon systemically absorbed in a rapid manner for either local lungor systemic treatment. An understanding of the precise mechanism,however, is not necessary for practicing the present invention asdescribed herein.

[0054] The aerosolized pharmaceutical-based dry powders of thisinvention are particularly useful in place of parenteral delivery. Thus,the methods and compositions of the present invention will beparticularly valuable in chronic treatment protocols where a patient canself-medicate. The patient can achieve a desired dosage by inhaling anappropriate amount of drug, as just described. The efficiency ofsystemic delivery via the method as just described will typically be inthe range from about 15% to 50%.

[0055] Method for Aerosolizing the Powder.

[0056] Still another aspect of this invention is a device and method foraerosolizing a pharmaceutical-based dry powder composition thatcomprises a therapeutically effective amount of drug in combination witha pharmaceutically acceptable carrier, which method comprises dispersingan amount of the dry powder composition in a gas stream to form anaerosol and capturing the aerosol in a chamber having a mouthpiece forsubsequent inhalation by a patient.

[0057] A further detailed description of this method is found in pendingU.S. patent application Ser. Nos. 07/910,048 and 08/207,472, both ofwhich are incorporated herein by reference.

[0058] Preparing the Compositions.

[0059] Still another aspect of this invention is a method for preparinga dispersible pharmaceutical-based dry powder composition of thisinvention that comprises spray drying an aqueous mixture of the drug anda pharmaceutically acceptable carrier under conditions to provide arespirable dry powder composition.

[0060] Spray drying is a process in which a homogeneous aqueous mixtureof drug and the carrier is introduced via a nozzle (e.g., a two fluidnozzle), spinning disc or an equivalent device into a hot gas stream toatomize the solution to form fine droplets. The aqueous mixture may be asolution, suspension, slurry, or the like, but needs to be homogeneousto ensure uniform distribution of the components in the mixture andultimately the powdered composition. Preferably the aqueous mixture is asolution. The solvent, generally water, rapidly evaporates from thedroplets producing a fine dry powder having particles 1 to 5 μm indiameter. Surprisingly, the drug is not degraded when it is exposed tothe hot drying gas, and the powders can be prepared having sufficientpurity for pharmaceutical use. An acceptable purity is defined as lessthan 5% degradation products and contaminates, preferably less than 3%and most preferably less than 1%.

[0061] The spray drying is done under conditions that result in asubstantially amorphous powder of homogeneous constitution having aparticle size that is respirable, a low moisture content and flowcharacteristics that allow for ready aerosolization. Preferably theparticle size of the resulting powder is such that more than about 98%of the mass is in particles having a diameter of about 10 μm or lesswith about 90% of the mass being in particles having a diameter lessthan 5 μm. Alternatively, about 95% of the mass will have particles witha diameter of less than 10 μm with about 80% of the mass of theparticles having a diameter of less than 5 μm.

[0062] The solutions may then be sprayed dried in conventional spraydrying equipment from commercial suppliers, such as Buchi, Niro, YamatoChemical Co., Okawara Kakoki Co., and the like, resulting in asubstantially amorphous particulate product.

[0063] For the spraying process, spraying methods such as rotaryatomization, pressure atomization and two-fluid atomization can be used.Examples of the devices used in these processes include “Parubisu[phonetic rendering] Mini-Spray GA-32” and “Parubisu Spray Drier DL-41”,manufactured by Yamato Chemical Co. “Spray Drier CL-8,” “Spray DrierL-8,” “Spray Drier FL-12,” “Spray Drier FL-16” or “Spray Drier FL-20,”manufactured by Okawara Kakoki Co., can be used for the method ofspraying using a rotary-disk atomizer.

[0064] While no special restrictions are placed on the nozzle of theatomizer used in the process of spraying, it is recommended to use anozzle which can produce a spray-dried composition with a grain diametersuitable for nasal, pharyngeal or pulmonary administration. For example,nozzle types “1A,” “1,” “2A,” “2,” “3” and the like, manufactured byYamato Chemical Co., can be used for the above-mentioned spray-drier,manufactured by the same company. In addition, disk types “MC-50,”“MC-65” or “MC-85,” manufactured by Okawara Kakoki Co., can be used asrotary disks of the spray-drier atomizer, manufactured by the samecompany.

[0065] While no particular restrictions are placed on the gas used todry the sprayed material, it is recommended to use air, nitrogen gas oran inert gas. The temperature of the inlet of the gas used to dry thesprayed materials is such that it does not cause heat deactivation ofthe sprayed material. The range of temperatures may vary between about50° C. to about 200° C., preferably between about 50° C. and 100° C. Thetemperature of the outlet gas used to dry the sprayed material may varybetween about 0° C. and about 150°, preferably between 0° C. and 90° C.,and even more preferably between 0° C. and 60° C. The fact that inletand outlet temperatures above about 55° C. can be used is surprising inview of the fact that most macromolecule-based drugs deactivate at thattemperature, with nearly complete deactivation occurring at about 70° C.

[0066] The dispersible pharmaceutical-based dry powders of the presentinvention may optionally be combined with pharmaceutical carriers orexcipients which are suitable for respiratory and pulmonaryadministration. Such carriers may serve simply as bulking agents when itis desired to reduce the pharmaceutical concentration in the powderwhich is being delivered to a patient, but may also serve to enhance thestability of the compositions and to improve the dispersibility of thepowder within a powder dispersion device in order to provide moreefficient and reproducible delivery of the powder and to improvehandling characteristics such as flowability and consistency tofacilitate manufacturing and powder filling.

[0067] Such carrier materials may be combined with the drug prior tospray drying, i.e., by adding the carrier material to the purified bulksolution. In that way, the carrier particles will be formedsimultaneously with the drug particles to produce a homogeneous powder.Alternatively, the carriers may be separately prepared in a dry powderform and combined with the dry powder drug by blending. The powdercarriers will usually be crystalline (to avoid water absorption), butmight in some cases be amorphous or mixtures of crystalline andamorphous. The size of the carrier particles may be selected to improvethe flowability of the drug powder, typically being in the range from 25μm to 100 μm. A preferred carrier material is crystalline lactose havinga size in the above-stated range.

[0068] Alternatively, dry powder compositions may be prepared by otherprocesses such as lyophilization and jet milling as disclosed inInternational Patent Publication No. WO 91/16038, the disclosure ofwhich is hereby incorporated by reference. TABLE 1 DRUG INDICATIONSSELECTED MACROMOLECULE DRUGS FOR SYSTEMIC APPLICATIONS CalcitoninOsteoporosis Prophylaxis Paget's Disease Hypercalcemia ErythropoietinAnemia Factor IX Hemophilia Granulocyte Colony Stimulating FactorNeutropenia (G-CSF) Granulocyte Macrophage Colony Stimulating BoneMarrow Engraftment/Transplant Failure Factor (GM-CSF) Growth HormoneShort stature Renal Failure Heparin Blood Clotting Heparin (LowMolecular Weight) Blood Clotting Insulin Type I and Type II DiabetesInterferon Alpha Hepatitus B and C Hairy Cell Leukemia Kaposi's SarcomaInterferon Beta Multiple Sclerosis Interferon Gamma ChronicGranulomatous Disease Interleukin-2 Renal Cancer Luteinizing HormoneReleasing Hormone Prostate Cancer (LHRH) Endometriosis SomatostatinAnalog Gastrointestinal Cancers Vasopressin Analog Diabetes InsipidusBed Wetting FSH Fertility Amylin Type I Diabetes Ciliary NeurotrophicFactor Lou Gehrig's Disease Growth Hormone Releasing Factor ShortStature Insulin-Like Growth Factor Osteoporosis Nutritional SupportInsulinotropin Type II Diabetes Interferon Beta Hepatitus B and CInterferon Gamma Rheumatoid Arthritus Interleukin-1 Receptor AntagonistRheumatoid Arthritus Interleukin-3 Adjuvant to ChemotherapyInterleukin-4 Immunodeficiency Disease Interleukin-6 ThrombocytopeniaMacrophage Colony Stimulating Fungal Disease Factor (M-CSF) CancerHypercholesterolemia Nerve Growth Factor Peripheral NeuropathiesParathyroid Hormone Osteoporosis Somatostatin Analog RefractoryDiarrheas Thymosin Alpha 1 Hepatitus B and C IIb/IIIa Inhibitor UnstableAngina Alpha-1 Antitrypsin Cystic Fibrosis Anti-RSV Antibody RespiratorySyncytial Virus Cystic Fibrosis Transmembrane Cystic Fibrosis Regulator(CFTR) Gene Deoxyribonuclase (DNase) Chronic Bronchitus Heparin AsthmaBactericidal/Permeability Adult Respiratory Distress Syndrome (ARDS)Increasing Protein Anti-CMV Antibody Cytomegalovirus Interleukin-1Receptor Asthma SELECTED NON-MACROMOLECULE DRUGS FOR SYSTEMIC AND LOCALLUNG APPLICATIONS Pentamidine isethiouate pneumocystis carini pneumoniaAlbuterol Sulfate Broncospasm Metaproterenol Sulfate Bronchial asthmaBeclomethasone Diprepionate Trimcinoline acetomide Budesonide acetonideIpratropium bromide Flunisolide Cromolyn sodium Ergotamine tartrateMigraines

[0069] The following examples are offered by way of illustration and notlimitation.

EXPERIMENTAL

[0070] According to the subject invention, the following dispersible drypowder formulations were prepared as described. All compositionsproduced according to the present invention meet the strictspecifications for content and purity required of pharmaceuticalproducts.

Example I 20.0% Insulin Formulation for Pulmonary Delivery

[0071] A. Formulation.

[0072] Bulk crystalline human zinc insulin was obtained from Eli Lillyand Company, Indianapolis, Ind. A 20% insulin formulation was achievedby combining 1.5 mg insulin per 1.0 mL deionized water with 4.96 mg/mLUSP mannitol and 1.04 mg/mL citrate buffer (sodium citrate dihydrate USPand citric acid monohydrate USP) for a total solids concentration of 7.5mg/mL at pH 6.7_(±)0.3.

[0073] B. Spray Drying.

[0074] A dry powder of the 20% insulin formulation described above wasproduced by spray drying the aqueous mixture using a Buchi LaboratorySpray Dryer under the following conditions: Temperature of aqueousmixture 2-8° C. Inlet temperature 120-122° C. Feed rate 5.3 mL/minOutlet temperature 80-81° C.

[0075] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at <80° C. for about 10 minutes by slowly decreasing theinlet temperature to provide a secondary drying.

[0076] C. Characterization.

[0077] The above 20% insulin dry powder composition contained 66.1%mannitol and 13.9% citrate. The composition was found to contain 1.1 to2.0% moisture as measured by a coulombic Karl Fischer method using aMitsubishi CA-06 Moisture Meter.

[0078] The particle size distribution of the composition was measured byliquid centrifugal sedimentation in a Horiba CAPA-700 Particle SizeAnalyzer following dispersion of the powder on Sedisperse A-11(Micrometrics, Norcross, Ga.) and was determined to be 1.3 μm to 1.5 μmMMD

[0079] The delivered dose of the insulin powder composition was measuredby collecting the aerosol powder produced by a dry powder dispersiondevice, similar to devices described in co-pending U.S. application Ser.Nos. 07/910,048; 08/313,707; 08/309,691 and PCT/US92/05621, thedisclosures of which are hereby incorporated by reference, on a filterplaced over the device mouthpiece. The delivered dose of the insulinpowder composition was determined to be 563±16 μg or 60 to 64% of thetotal powder (5.0 mg) loaded into the device.

[0080] The aerosol particle size distribution, measured using a cascadeimpactor (California Measurements IMPAQ-6), was determined to be 2.0 μmMMAD, with 86% to 90% of the particles <5.0 μm in diameter.

[0081] The insulin content of the powder, measured by reverse phase HPLC(rpHPLC) was determined to be 197 μg/mg powder, accounting for 99% ofthe expected insulin. No degradation peaks were detected in thechromatogram.

Example II 5.0% Parathyroid Hormone Formulation for Pulmonary Delivery

[0082] A. Formulation.

[0083] Bulk 34 amino acid active fragment of parathyroid hormone, PTH(1-34), was obtained from BACHEM CALIFORNIA, Torrance, Calif. A 5.0% PTH(1-34) formulation was achieved by combining 0.375 mg PTH (1-34) per 1.0mL deionized water with 6.06 mg/mL mannitol USP and 1.04 mg/mL citratebuffer (sodium citrate dihydrate USP and citric acid monohydrate USP)for a total solids concentration of 7.48 mg/mL at pH 6.3.

[0084] B. Spray Drying.

[0085] A dry powder of the 5.0% PTH (1-34) formulation described abovewas produced by spray drying the aqueous mixture using a BuchiLaboratory Spray Dryer under the following conditions: Temperature ofaqueous mixture 2-8° C. Inlet temperature 122-124° C. Feed rate 5.2mL/min Outlet temperature 73-74° C.

[0086] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at <80° C. for about 5 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0087] C. Characterization.

[0088] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0089] The above 5.0% PTH (1-34) dry powder composition contained 81.0%mannitol and 13.9% citrate. The formulation contained 0.5% moisture.

[0090] The particle size distribution of the composition was determinedto be 2.4 μm and 2.7 μm MMD in separate measurements.

[0091] The delivered dose of the PTH (1-34) powder was determined to be161 μg or 64.5% and 175 μg or 69.2% in separate measurements.

[0092] The PTH (1-34) content of the powder, measured by rpHPLC wasdetermined to be 48.5 μg/mg powder, accounting for 97% of the expectedvalue. No degradation peaks were detected in the chromatogram.

Example III 0.7% Interleukin-1 Receptor Formulation for PulmonaryDelivery

[0093] A. Formulation.

[0094] Bulk interleukin-1 receptor, IL-1 receptor, was obtained fromImmunex Corporation, Seattle, Wash. A 0.7% IL-1 receptor formulation wasachieved by combining 0.053 mg IL-1 receptor per 1.0 mL deionized waterwith 7.07 mg/mL raffinose (Pfanstiehl, Waukegan, Ill.) and 0.373 mg/mLTris buffer at pH 7.18.

[0095] B. Spray Drying.

[0096] A dry powder of the 0.7% IL-1 receptor formulation describedabove was produced by spray drying the aqueous mixture using a BuchiLaboratory Spray Dryer under the following conditions: Temperature ofaqueous mixture 2-8° C. Inlet temperature 135-137° C. Feed rate 4.9mL/min Outlet temperature 92-93° C.

[0097] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at 90° C. for about 15 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0098] C. Characterization.

[0099] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0100] The above 0.7% IL-1 receptor dry powder composition contained94.3% raffinose and 5.0% Tris. The formulation contained 1.84±0.25%moisture.

[0101] The particle size distribution of the composition was determinedto be 1.95 μm MMD with 100% of the particles <5.0 μm.

[0102] The delivered dose of the IL-1 receptor powder was determined tobe 22.3±2.0 μg or 53.4±4.7%.

[0103] The aerosol particle size distribution, was determined to be 3.2μm MMAD, with 77% of the particles <5.0 μm in diameter.

[0104] The IL-1 receptor content of the powder as measured by rpBPLC wasdetermined to be 8.4 μg/mg, accounting for 120% of the expected IL-1receptor. No degradation peaks were detected in the chromatogram.

Example IV 5.0% Interleukin-1 Receptor Formulation for PulmonaryDelivery

[0105] A. Formulation.

[0106] Bulk interleukin-1 receptor, IL-1 receptor, was obtained fromImmunex Corporation, Seattle, Wash. A 5.0% IL-1 receptor formulation wasacheived by combining 0.375 mg IL-1 receptor per 1.0 mL deionized waterwith 6.77 mg/mL raffinose and 0.351 mg/mL Tris buffer at pH 7.35.

[0107] B. Spray Drying.

[0108] A dry powder of the 5.0% IL-1 receptor formulation describedabove was produced by spray drying the aqueous mixture using a BuchiLaboratory Spray Dryer under the following conditions: Temperature ofaqueous mixture 2-8° C. Inlet temperature 138° C. Feed rate 4.9 mL/minOutlet temperature 91° C.

[0109] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at 90° C. for about 15 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0110] C. Characterization.

[0111] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0112] The above 5.0% IL-1 receptor dry powder composition contained90.3% raffinose and 4.7% Tris. The formulation contained 1.75±0.26%moisture.

[0113] The particle size distribution of the composition was determinedto be 2.74 μm MMD with 97% of the particles <5.0 μm.

[0114] The delivered dose of the IL-1 receptor powder was determined tobe 123.4±24.5 μg or 49.3±9.8%.

[0115] The aerosol particle size distribution, was determined to be 4.1μm MMAD, with 64% of the particles <5.0 μm in diameter.

[0116] The IL-1 receptor content of the powder as measured by rpHPLC wasdetermined to be 52.7±1.8 μg/mg, accounting for 105% of the expectedIL-1 receptor. No degradation peaks were detected in the chromatogram.

Example V 26.7% Human Calcitonin Formulation for Pulmonary Delivery

[0117] A. Formulation.

[0118] Bulk human calcitonin was obtained from Ciba-Geigy. A 26.7% humancalcitonin formulation was acheived by combining 1.9 mg human calcitoninper 1.0 mL deionized water with 4.3 mg/mL mannitol and 0.9 mg/mL citratebuffer at pH 3.85.

[0119] B. Spray Drying.

[0120] A dry powder of the 26.7% human calcitonin formulation describedabove was produced by spray drying the aqueous mixture using a BuchiLaboratory Spray Dryer under the following conditions: Temperature ofaqueous mixture 4° C. Inlet temperature 119° C. Feed rate 5.5 mL/minOutlet temperature 78° C. Atomizer coolant temperature 0-5° C. Cyclonecoolant temperature 25-30° C.

[0121] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at 80° C. for about 10 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0122] C. Characterization.

[0123] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0124] The above 26.7% human calcitonin dry powder composition contained60% mannitol and 13.3% citrate. The formulation contained 0.71%moisture.

[0125] The particle size distribution of the composition was determinedto be 1.33±0.63 μm MMD.

[0126] The delivered dose of the human calcitonin powder was determinedto be 76.8±6.7%.

[0127] The human calcitonin content of the powder as measured by rpHPLCwas determined to be 272.0 μg/mg, accounting for 102±1.7% of theexpected human calcitonin. No degradation peaks were detected in thechromatogram.

Example VI 90% Alpha-1 Antitrypsin Formulation for Pulmonary Delivery

[0128] A. Formulation.

[0129] Bulk alpha-1 antitrypsin, A1A, was obtained from ArmourPharmaceutical Company, Kankakee, Ill. A 90% A1A formulation wasacheived by combining 4.89 mg A1A per 1.0 mL deionized water with 0.54mg/mL citrate buffer at pH 6.0.

[0130] B. Spray Drying.

[0131] A dry powder of the 90% A1A formulation described above wasproduced by spray drying the aqueous mixture using a Buchi LaboratorySpray Dryer under the following conditions: Temperature of aqueousmixture 4° C. Inlet temperature 98-101° C. Feed rate 5.0 mL/min Outlettemperature 65° C. Atomizer coolant temperature 2-8° C. Cyclone coolanttemperature 30° C.

[0132] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at 69° C. for about 10 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0133] C. Characterization.

[0134] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0135] The above 90% A1A dry powder composition contained 10.0% citrate.The formulation contained 4.79% moisture.

[0136] The particle size distribution of the composition was determinedto be 1.71±0.87 μm MMD.

[0137] The delivered dose of the 90% A1A powder was determined to be67.0±5.0%.

[0138] The aerosol particle size distribution, was determined to be 1.0μm MMAD, with 90% of the particles <5.0 μm in diameter.

[0139] The A1A content of the powder,as measured by rpHPLC wasdetermined to be 80% of the expected value. No degradation peaks weredetected in the chromatogram. The activity after spray drying wasdetermined to be 74±1%

Example VII 0.3% Beta Interferon Formulation for Pulmonary DeliveryContaining Human Serum Albumin

[0140] A. Formulation.

[0141] Bulk beta interferon, IFN-β, was obtained from Toray Industries,Inc., Tokyo, Japan. A 0.3% IFN-β formulation was acheived by combining0.025 mg IFN-β per 1.0 mL deionized water with 5.54 mg/mL human serumalbuman (HSA), 2.3 mg/mL citrate buffer and 0.345 mg/mL of NaCl at pH4.5.

[0142] B. Spray Drying.

[0143] A dry powder of the 0.3% IFN-β formulation described above wasproduced by spray drying the aqueous mixture using a Buchi LaboratorySpray Dryer under the following conditions: Temperature of aqueousmixture 2-8° C. Inlet temperature 93° C. Feed rate 2.7 mL/min Outlettemperature 62° C.

[0144] C. Characterization.

[0145] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0146] The above 0.3% IFN-β dry powder composition contained 66.0% HSA,27.4% citrate, 4.1% NaCl. The formulation contained 4.22% moisture.

[0147] The particle size distribution of the composition was determinedto be 1.62 μm MMD with 94.8% of the particles <5 μm.

[0148] The delivered dose of the 0.3% IFN-β powder was determined to be9.9 μg/mg or 66.0±4.0%.

[0149] The aerosol particle size distribution, was determined to be 2.0μm MMAD, with 85% of the particles <5.0 μm in diameter.

[0150] The IFN-β activity of the powder as measured by IFN-β enzymeimmunoassay (Toray-Fuji Bionics) and was determined to be 109±8% of theexpected activity.

Example VIII 0.3% Beta Interferon Formulation for Pulmonary DeliveryContaining Raffinose

[0151] A. Formulation.

[0152] Bulk beta interferon, IFN-β, was obtained from Toray Industries,Inc., Tokyo, Japan. A 0.3% IFN-β formulation was achieved by combining0.025 mg IFN-β per 1.0 mL deionized water with 4.7 mg/mL raffinose, 1.0mg/mL human serum albuman (HSA), 2.3 mg/mL citrate buffer and 0.3 mg/mLof NaCl at pH 4.5.

[0153] B. Spray Drying.

[0154] A dry powder of the 0.3% IFN-β formulation described above wasproduced by spray drying the aqueous mixture using a Buchi LaboratorySpray Dryer under the following conditions: Temperature of aqueousmixture 2-8° C. Inlet temperature 145° C. Feed rate 5.0 mL/min Outlettemperature 87° C.

[0155] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at 97° C. for about 5 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0156] C. Characterization.

[0157] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0158] The above 0.3% IFN-β dry powder composition contained 56.4%raffinose, 11.9% HSA, 27.4% citrate, 3.5% NaCl. The formulationcontained 0.69% moisture.

[0159] The particle size distribution of the composition was determinedto be 2.06 μm MMD with 88.9% of the particles <5 μm .

[0160] The delivered dose of the 0.3% IFN-β powder was determined to be10.2 μg/mg or 68.0±2.0%.

[0161] The aerosol particle size distribution, was determined to be 2.5μm MMAD, with 84% of the particles <5.0 μm in diameter.

[0162] The IFN-β activity of the powder as measured by INF-β enzymeimmunoassay (Toray-Fuji Bionics) and was determined to be 109±8% of theexpected activity.

Example IX 93% Low Molecular Weight Heparin Formulation for PulmonaryDelivery

[0163] A. Formulation.

[0164] Bulk low molecular weight heparin sodium salt (Av. Mol. Wt.:Approx. 6000) from porcine intestinal mucosa, heparin (LMW), wasobtained from Sigma Chemical, St. Louis, Mo. A 93% heparin (LMW)formulation was achieved by combining 6.9 mg heparin (LMW) per 1.0 mLdeionized water with 0.5 mg/mL HSA at pH 6.9.

[0165] B. Spray Drying.

[0166] A dry powder of the 93% heparin (LMW) formulation described abovewas produced by spray drying the aqueous mixture using a BuchiLaboratory Spray Dryer under the following conditions: Temperature ofaqueous mixture 2-8° C. Inlet temperature 140° C. Feed rate 3.8 mL/minOutlet temperature 85° C. Atomizer coolant temperature 2-8° C. Cyclonecoolant temperature 20° C.

[0167] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at 80° C. for about 10 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0168] C. Characterization.

[0169] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0170] The above 93% heparin (LMW) dry powder composition contained 7.0%HSA.

[0171] The delivered dose of the 93% heparin (LMW) powder was determinedto be 60.0±1.0%.

[0172] The aerosol particle size distribution, was determined to be 3.5μm MMAD, with 70% of the particles <5.0 μm in diameter.

Example X 97% Unfractionated Heparin Formulation for Pulmonary Delivery

[0173] A. Formulation.

[0174] Bulk unfractionated heparin sodium salt from porcine intestinalmucosa, heparin, was obtained from Sigma Chemical, St. Louis, Mo. A 97%heparin formulation was achieved by combining 7.0 mg heparin per 1.0 mLdeionized water with 0.25 mg/mL HSA at pH 6.55.

[0175] B. Spray Drying.

[0176] A dry powder of the 97% heparin formulation described above wasproduced by spray drying the aqueous mixture using a Buchi LaboratorySpray Dryer under the following conditions: Temperature of aqueousmixture 2-8° C. Inlet temperature 150° C. Feed rate 4.0 mL/min Outlettemperature 85° C. Atomizer coolant temperature 2-8° C. Cyclone coolanttemperature 20° C.

[0177] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at 80° C. for about 10 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0178] C. Characterization.

[0179] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0180] The above 97% heparin dry powder composition contained 3.0% HSA.The formulation contained 5.11% moisture.

[0181] The particle size distribution of the composition was determinedto be 2.0 to 2.5 μm MMD.

[0182] The delivered dose of the 97% heparin powder was determined to be79.0±6.0%.

[0183] The aerosol particle size distribution, was determined to be 3.2μm MMAD, with 70% of the particles <5.0 μm in diameter.

Example XI Lipid Vector Gene Formulation for Pulmonary Delivery

[0184] A. Formulation.

[0185] Bulk pCMVβ DNA:Lipid vector as described in U.S. application Ser.No. 08/417,507 filed Apr. 14, 1995 entitled, “COMPOSITIONS AND METHODSFOR NUCLEIC ACID DELIVERY TO THE LUNG”, the disclosure of which ishereby incorporated by reference, was obtained from Genzyme Corporation,Cambridge, Mass. A 0.71% DNA:Lipid vector formulation was acheived bycombining 0.005:0.03 mg DNA:Lipid vector per 1.0 mL deionized water with5.3 mg/mL glycine (J. T. Baker) 0.3 mg/mL HSA at pH 6.4.

[0186] B. Spray Drying.

[0187] A dry powder of the DNA:Lipid vector formulation described abovewas produced by spray drying the aqueous mixture using a BuchiLaboratory Spray Dryer under the following conditions: Temperature ofaqueous mixture 2-8° C. Inlet temperature 120° C. Feed rate 3.8 mL/minOutlet temperature 71° C. Atomizer coolant temperature 2-8° C. Cyclonecoolant temperature 2-8° C.

[0188] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at 65° C. for about 5 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0189] C. Characterization.

[0190] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0191] The above 0.71% DNA:Lipid vector dry powder composition contained93.97% glycine, and 5.32% HSA.

[0192] The particle size distribution of the composition was determinedto be 2.0 μm MMD.

[0193] The delivered dose of the powder was determined to be 64.0±1.0%.

[0194] The aerosol particle size distribution, was determined to be 2.4μm MMAD, with 75% of the particles <5.0 μm in diameter.

[0195] Activity after spray drying was determined to be 160% of theexpected value.

Example XII Adenoviral Vector Gene Formulation for Pulmonary Delivery

[0196] A. Formulation.

[0197] Bulk pCMVβ DNA:Adenovirous vector as described in U.S.application Ser. No. 08/417,507 filed Apr. 14, 1995 entitled,“COMPOSITIONS AND METHODS FOR NUCLEIC ACID DELIVERY TO THE LUNG”, thedisclosure of which is hereby incorporated by reference, was obtainedfrom Genzyme Corporation, Cambridge, Mass. A DNA:adenovirous vectorformulation was achieved by combining 108 PFU//mL DNA:Lipid vector per1.0 mL deionized water with 6.1 mg/mL glycine J. T. Baker) 2.5 mg/mLHSA, 1.9 mg/mL phosphate buffer at pH 7.4.

[0198] B. Spray Drying.

[0199] A dry powder of the DNA:Lipid vector formulation described abovewas produced by spray drying the aqueous mixture using a BuchiLaboratory Spray Dryer under the following conditions: Temperature ofaqueous mixture 2-8° C. Inlet temperature 105° C. Feed rate 2.9 mL/minOutlet temperature 72° C. Atomizer coolant temperature 2-8° C. Cyclonecoolant temperature 20° C.

[0200] Once the aqueous mixture was consumed, the outlet temperature wasmaintained at 70° C. for about 10 minutes by slowly decreasing the inlettemperature to provide a secondary drying.

[0201] C. Characterization.

[0202] The following characterization of the dry powder formulationdescribed above was carried out using the methods described in Example Iunless indicated otherwise.

[0203] The above DNA:adenovirous vector dry powder composition contained58% glycine, and 24% HSA and 18% phosphate buffer.

[0204] The particle size distribution of the composition was determinedto be 2.3 μm MMD.

[0205] The delivered dose of the powder was determined to be 51.0±1.0%.

[0206] The aerosol particle size distribution, was determined to be 1.8μm MMAD, with 80% of the particles <5.0 μm in diameter.

[0207] Activity after spray drying was determined to be 76% of theexpected value.

[0208] All publications and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication or patent application was specificallyand individually indicated to be incorporated by reference.

[0209] The invention now being fully described, it will be apparent toone of ordinary skill in the art that many changes and modifications canbe made thereto without departing from the spirit or scope of theappended claims.

The subject matter claimed is:
 1. An aerosolizable, spray-dried powderformulation for pulmonary delivery comprising a therapeuticallyeffective amount of FSH.
 2. The formulation of claim 1, containing lessthan 5% FSH degradation products.
 3. The formulation of claim 1, furthercomprising a pharmaceutically acceptable excipient.
 4. The formulationof claim 3, wherein said excipient is selected from the group consistingof carbohydrates, amino acids, polypeptides, buffers, and salts.
 5. Theformulation of claim 4, wherein said excipient is a carbohydrateselected from the group consisting of galactose, mannose, sorbose,lactose, trehalose, cyclodextrin, raffinose, maltodextrins, dextrans,mannitol, and xylitol.
 6. The formulation of claim 4, wherein saidexcipient is an amino acid.
 7. The formulation of claim 6, wherein saidamino acid is selected from the group consisting of alanine, glycine,tryptophan, tyrosine, leucine, and phenylalanine.
 8. The formulation ofclaim 6, wherein said amino acid is a hydrophobic amino acid.
 9. Theformulation of claim 8, wherein said amino acid is effective to increasethe dispersibility of the formulation.
 10. The formulation of claim 4,wherein said excipient comprises HSA.
 11. The formulation of claim 4,wherein said excipient comprises a buffer.
 12. The formulation of claim4, comprising from about 0.05 to about 99 percent by weight FSH.
 13. Theformulation of claim 1, wherein said powder comprises particles with anaverage particle of less than 10 microns MMD.
 14. The formulation ofclaim 13, comprising particles sized from about 1.0-5.0 microns MMD. 15.The formulation of claim 1, wherein said powder comprises particlessized from about 1.0-5.0 microns MMAD.
 16. The formulation of claim 1,further characterized by a delivered dose of greater than about 30%. 17.The formulation of claim 1, aerosolizable in a dry powder inhaler.
 18. Amethod of treating a disease state responsive to treatment by FSH, saidmethod comprising administering by inhalation to a subject in needthereof the formulation of claim 1 in aerosolized form.
 19. The methodof claim 18, wherein said administering step comprises dispersing saidpowder formulation in a gas stream to form an aerosol and inhaling. 20.A method for preparing a FSH dry powder composition suitable forpulmonary delivery, said method comprising spray drying an aqueousmixture comprising FSH under conditions effective to provide arespirable, spray dried FSH powder.
 21. The method of claim 20, whereinthe powder formed in said spray drying step contains less than 5% FSHdegradation products.
 22. The method of claim 20, wherein said aqueousmixture further comprises a pharmaceutically acceptable excipient. 23.The method of claim 20, wherein 98% or more of the mass of the spraydried powder comprises particles having a diameter of 10 microns orless.
 24. The method of claim 20, wherein 90% or more of the mass of thespray dried powder comprises particles having a diameter of 5 microns orless.
 25. A respirable powder produced by the method of claim 20.